Modified polyamine polymers for delivery of biomolecules into cells

Abstract

Provided herein are compounds, compositions, and methods for delivering biomolecules to cells. In particular, the present disclosure provides modified polyamine polymers, including polyamine polymers having fluorinated substituents.

1 . A compound or a salt thereof, the compound comprising:
 a polyethyleneimine polymer; and   a plurality of substituents bound to amino groups of the polyethyleneimine polymer, wherein each substituent independently has a formula (I):
   —X—(CH 2 ) n —Z   (I),
 
   wherein:   X is a bond or —C(O)—O—;   n is 0, 1, 2, 3, 4, or 5; and   Z is selected from a haloalkyl group, an aryl group, a substituted aryl group, a heteroaryl group, and a substituted heteroaryl group.   
     
     
         2 . The compound of  claim 1 , or a salt thereof, wherein the polyethyleneimine polymer has a weight average molecular weight of about 500 Da to about 250000 Da. 
     
     
         3 .- 4 . (canceled) 
     
     
         5 . The compound of  claim 1 , or a salt thereof, wherein the polyethyleneimine polymer is a branched polyethyleneimine polymer. 
     
     
         6 . The compound of  claim 1 , or a salt thereof, wherein the polyethyleneimine polymer is a linear polyethyleneimine polymer. 
     
     
         7 . The compound of  claim 1 , or a salt thereof, wherein Z is a haloalkyl group having the following formula:
 —(CF 2 ) m —CF 3 , wherein m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.   
     
     
         8 . The compound of  claim 1 , or a salt thereof, wherein Z is a pentafluorophenyl group or an unsubstituted pyridyl group. 
     
     
         9 . (canceled) 
     
     
         10 . The compound of  claim 1 , or a salt thereof, wherein:
 X is —C(O)O—; and   n is 1 or 2.   
     
     
         11 . (canceled) 
     
     
         12 . The compound of  claim 1 , or a salt thereof, wherein
 X is a bond; and   n is 1 or 2.   
     
     
         13 . The compound of  claim 1 , or a salt thereof, wherein about 0.1 mol % to about 60 mol % of the amino groups of the polyethyleneimine polymer are bound to a substituent of formula (I). 
     
     
         14 . (canceled) 
     
     
         15 . The compound of  claim 13 , or a salt thereof, wherein about 8 mol % to about 40 mol % of the amino groups of the polyethyleneimine polymer are bound to a substituent of formula (I). 
     
     
         16 . A compound or a salt thereof, the compound comprising:
 a poly(amidoamine) dendrimer; and   a plurality of substituents bound to amino groups of the poly(amidoamine) dendrimer, wherein each substituent independently has a formula (I):
   —X—(CH 2 ) n —Z   (I),
 
   wherein:   X is a bond or —C(O)—O—;   n is 0, 1 or 2; and   Z is selected from a haloalkyl group, an aryl group, a substituted aryl, a heteroaryl group, and a substituted heteroaryl group.   
     
     
         17 - 27 . (canceled) 
     
     
         28 . A method of delivering a biomolecule to a cell, comprising:
 contacting the cell with an effective amount of a compound of  claim 1 , or a salt thereof; and   contacting the cell with the biomolecule.   
     
     
         29 . (canceled) 
     
     
         30 . The method of  claim 28 , wherein the biomolecule is at least one of a deoxyribonucleic acid (DNA) molecule, a ribonucleic acid (RNA) molecule, a peptide, a polypeptide, a protein, or any combinations or derivatives thereof. 
     
     
         31 .- 32 . (canceled) 
     
     
         33 . The method of  claim 28 , wherein the biomolecule comprises a polypeptide sequence of SEQ ID NO: 4. 
     
     
         34 . The method of  claim 28 , wherein the biomolecule is a ribonucleoprotein complex comprising a Cas9 protein. 
     
     
         35 .- 36 . (canceled) 
     
     
         37 . A kit comprising a compound of  claim 1 , or a salt thereof. 
     
     
         38 . (canceled) 
     
     
         39 . The kit of  claim 37 , further comprising at least one of a DNA molecule, an RNA molecule, a peptide, a polypeptide, a protein, or any combinations or derivatives thereof. 
     
     
         40 .- 41 . (canceled) 
     
     
         42 . The kit of  claim 37 , further comprising instructions for using the compound or the salt thereof for transfection of a biomolecule. 
     
     
         43 . A method for altering a sequence of an endogenous protein in a cell, the method comprising:
 assembling a ribonucleoprotein (RNP) complex comprising a Cas9 protein, a donor DNA template, and a guide RNA; and   delivering the RNP complex into a cell using a compound of  claim 1 .   
     
     
         44 . A method for tagging an endogenous protein in a cell, the method comprising:
 assembling a ribonucleoprotein (RNP) complex comprising a Cas9 protein, a donor DNA template, and a guide RNA, wherein the donor DNA template comprises a sequence encoding a peptide or polypeptide tag sequence; and   delivering the RNP complex into a cell using a compound of  claim 1 .   
     
     
         45 .- 46 . (canceled)